The purpose of this project is to determine the function of thiamine in nerve conduction. The specific experiments will be in the form of three different approaches. The first approach will be to isolate acetyl choline receptor sites from electric eel tissue and to measure their binding constant with thiamine and thiamine derivatives. The positive charge on the thiamine molecule makes thiamine a likely candidate for a competitive inhibitor of acetyl choline for the receptor site, particularly since acetyl cholinesterase is inhibited by thiamine. We plan to look for cooperativity effects in the thiamine binding if it does bind. Our second approach will be to make affinity columns with the thiamine molecule attached to the column in order to isolate thiamine binding proteins. We will use electric eel tissue and mouse brain for these experiments. A third approach will be to modify the thiamine molecule so that it will react with the binding protein in the nerve. Our first experiments have been to prepare O-bromoacetyl and O-chloroacetyl thiamine. We have found that these compounds are anesthetics in mice. We plan to continue this work by preparing other O-acyl derivatives of thiamine and testing their activities as anesthetics. BIBLIOGRAPHIC REFERENCE: Waleh, A. and Ingraham, L.L. (1976). Electron transport in cytochrome c. Proceedings of the Eighth Jerusalem Conference on Quantum Chemistry and Biochemistry (B. Pullman, ed.), pp. 505-516, Reidel Publ., Boston.